Device for automated bioreactor sampling

Abstract

The invention relates to a device for automated bioreactor sampling connected to a reactor comprising a sampler, at least one tube fluidly connecting said reactor with said sampler and a pump for pumping fluids from said reactor to said sampler and vice versa. Particularly simple and essentially dead volume-free sampling can be achieved by providing means for flushing at least parts of the tube with gas/sterile air between individual sampling steps. The invention furthermore relates to a method of operation of such a device.

1 - 14 . (canceled)  
     
     
         15 . A device for automated bioreactor sampling to be connected to a reactor, comprising, a sampler, at least one tube fluidly connecting said reactor with said sampler and a pump for pumping fluids from said reactor to said sampler and vice versa, wherein means are provided for flushing at least parts of the tube with gas between individual sampling steps.  
     
     
         16 . The device according to  claim 15 , wherein said gas is flushing the tube such that fluids in the tube are pushed back into the reactor after the sampling and/or that said gas is flushing the tube such that fluids in the tube are pushed into the sampler for finishing the sampling.  
     
     
         17 . The device according to  claim 15 , wherein the pump is connected with or integrated into the tube fluidly connecting said reactor with said sampler.  
     
     
         18 . The device according to  claim 17 , wherein at least one of the tubes is a flexible tube such as a silicone tube.  
     
     
         19 . The device according to  claim 17 , wherein the flexible tube is a silicone tube.  
     
     
         20 . The device according to  claim 17 , wherein the pump is a peristaltic pump.  
     
     
         21 . The device according to  claim 17 , wherein there is provided a biomass breaker which on its upper part is connected with the reactor by a first tube and which on the bottom is connected with the sampler by a second tube, wherein the flushing gas is introduced into the system on the upper part of the biomass breaker, and wherein the second tube is a flexible tube and the pump is a peristaltic pump acting upon said flexible tube.  
     
     
         22 . The device according to  claim 21 , wherein means are provided for introducing rinse solution and/or inactivator solution into at least parts of the tubes between individual sampling steps.  
     
     
         23 . The device according to  claim 22 , wherein said solution is introduced into the system in the biomass breaker.  
     
     
         24 . The device according to  claim 22 , wherein there is provided at least one rinse/inactivator solution container connected with the biomass breaker by means of flexible tubes, wherein the flow of the solution is effected by a peristaltic pump and the flow is controlled by means of pinch valves.  
     
     
         25 . The device according to  claim 24 , wherein the peristaltic pump for the rinse/inactivator solution is driven by the same shaft as the peristaltic pump for the samples.  
     
     
         26 . The device according to  claim 24 , wherein a bypass is provided for recirculating the solution when it shall not be mixed with the samples.  
     
     
         27 . The device according to  claim 15 , wherein there is provided a waste receptacle for taking up undesired fluids, and in that parts of the tube can automatically be shifted from the sampler to this waste receptacle.  
     
     
         28 . The device according to  claim 15 , wherein the sampler is a fraction collector allowing controlled cooling of the sample containers.  
     
     
         29 . The device according to  claim 28 , wherein the sample containers are partially embedded in a block of material such as aluminium which can be cooled directly or indirectly by a cooling fluid provided by a flow cooler.  
     
     
         30 . The device according to  claim 20 , wherein the cooling fluid is circulated through internal channels of the block.  
     
     
         31 . The device according to  claim 15 , wherein a sterile filter, with a pore diameter in the range of 0.2 μm, and/or a wet separator are provided in the piping for the air for flushing.  
     
     
         32 . A process for sampling of a bioreactor using a device according to  claim 15 , the process comprising the step of sampling the bioreactor.  
     
     
         33 . The process according to  claim 32 , wherein 
 in a first step, the resting state between sampling, a robotic arm is positioned above drain that leads to a waste receptacle while at least the tube between reactor and biomass breaker is empty while the tube between reactor and drain is either empty or filled with rinse solution,    in a second step, the peristaltic pump is turned on to empty the tube of rinse solution, the pump is subsequently turned off while the robotic arm moves to position above the next empty test tube in the sampler, the pump is subsequently turned on again for a time period necessary to fill the tube with sample/broth from the reactor, the robotic arm subsequently returns to position above drain once pump turns off,    in a third step, the pinch valves are activated opening gas/air and rinse solution tubing, while closing bypass tubing, such that sterile air pushes sample broth back into reactor, while as soon as the pump is turned on, sterile rinse solution enters biomass breaker and travels through tubing to waste receptacle, and    in a fourth step, the peristaltic pump and rinse solution and bypass are turned off, while gas/air valve remains activated for a short time period to confirm that tubing from biomass breaker to reactor is empty, while as soon as the pump is turned off, the air flow rate through this tubing is augmented, the gas/air valve is then deactivated to block gas/air.    
     
     
         34 . The process according to  claim 32 , wherein: 
 in a first step, the resting state between sampling, a robotic arm is positioned above drain that leads to a waste receptacle while at least the tube between reactor and biomass breaker is empty while the tube between reactor and drain is either empty or filled with rinse solution,    in a second step, the robotic arm moves to position above the next empty test tube in the sampler, the pump is subsequently turned on for a time period necessary to fill the test tube with sample/broth from the reactor and, optionally, an inactivator solution like perchloric acid is pumped simultaneously and mixed with sample in biomass breaker,    in a third step, the pinch valves are activated opening air and bypass tubing, while closing inactivator solution tubing such that sterile air pushes sample broth back into reactor, with the pump on, air pushes remainder of sample/inactivator solution through tubing and into sample tube,    in a fourth step, the peristaltic pump and inactivator and bypass are turned off and the robotic arm returns to position above drain the gas/air valve remains activated for a short time period to confirm that tubing from biomass breaker to reactor is empty, while as soon as the pump is turned off, the airflow rate through this tubing is augmented and the gas/air valve is then deactivated to block gas/air.    
     
     
         35 . The process according to  claim 32 , wherein the fluid is kept in the sampler at a temperature below 2° C. and above freezing temperature.  
     
     
         36 . The process according to  claim 32 , wherein the fluid is kept in the sampler at a temperature in the range of 0.1° C.  
     
     
         37 . The process according to  claim 32 , wherein fluid can be cooled down to temperatures below 2° C. within <20 minutes.  
     
     
         38 . The process according to  claim 32 , wherein fluid can be cooled down to temperatures below 1° C. within about 10 minutes.